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Department of Medical Nutrition and Center for Biotechnology, Karolinska Institute, S-14186 Huddinge, Sweden
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ABSTRACT |
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 TOP ABSTRACT INTRODUCTIONRESULTS AND DISCUSSIONMATERIALS AND METHODSREFERENCES |
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(ER) and
activates transcription of reporter gene constructs containing
estrogen-response elements in transient transfections in response to
estradiol. Using a mammalian two-hybrid system, the formation of
heterodimers of the ERß and ER subtypes was demonstrated.
Furthermore, ERß and ER form heterodimeric complexes with retained
DNA-binding ability and specificity in vitro. In addition,
DNA binding by the ERß/ER heterodimer appears to be dependent on
both subtype proteins. Taken together these results suggest the
existence of two previously unrecognized pathways of estrogen
signaling; I, via ERß in cells exclusively expressing this subtype,
and II, via the formation of heterodimers in cells expressing both
receptor subtypes. |
INTRODUCTION |
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TOPABSTRACTINTRODUCTIONRESULTS AND DISCUSSIONMATERIALS AND METHODSREFERENCES |
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Steroid hormone receptors constitute a distinct subgroup within the nuclear receptor family (2), which includes receptors for glucocorticoids, mineralocorticoids, androgens, progestins, and estrogens [glucocorticoid receptor, mineralocorticoid receptor, androgen receptor, progesterone receptor, and estrogen receptor (ER), respectively]. In addition two orphan nuclear receptors, the ERR-1 and 2 (ER-related receptors) (3) have been referred to this group (4). The steroid hormone receptors bind as homodimers to palindromic DNA response elements (2). Another important feature of steroid hormone receptors is the interaction with the molecular chaperone hsp 90 (5).
Estrogens influence growth, differentiation, and function of many
target tissues, including tissues of the female and male reproductive
tract (6). Estrogens also play an important role in the maintenance of
bone mass and in the cardiovascular system where estrogens have certain
protective effects (7, 8). The ER-encoding cDNAs have been cloned from
several species (9, 10, 11, 12). Important examples of genes regulated by
estrogens are the PR, epidermal growth factor receptor, certain growth
factors (insulin-like growth factor-I, transforming growth factor-
and -ß) and several protooncogenes (c-fos,
c-myc, c-jun) (13). Loss of ER function has long
been postulated to be incompatible with life, and therefore the
successful generation of ER-deficient mice came as a surprise (14).
These mice are viable but display severe dysfunction of the
reproductive organs, and both sexes are sterile. The females have
hypoplastic uteri and hyperemic ovaries with no detectable corpora
lutea. The fact that disruption of the ER gene did not completely
eliminate the ability of small follicles to grow, as was evident from
the presence of secondary and antral follicles in the knock-out mouse
ovary, pointed to the possible existence of alternative ER mediating
the intraovarian effects of estradiol. In some tissues from the ER
knock-out mice residual binding of estradiol with an affinity and
specificity reminiscent of an ER protein could be measured (14, 15). We
have recently cloned a novel ER cDNA from rat prostate (16), which was
suggested to be named rat ERß subtype to distinguish it from the
previously cloned ER cDNA (consequently ER subtype). The rat ERß
protein was found to be highly homologous to the rat ER protein,
particularly in the DBD (>95% amino acid identity) and in the
C-terminal ligand-binding domain (55% amino acid identity). In
ligand-binding assays rat ERß binds estrogens with an affinity and
specificity resembling that of ER, and ERß is able to activate
transcription of an estrogen-response element containing reporter gene
construct (16, 17). In subsequent studies it was shown that ERß is
the primary ER subtype expressed in rat ovary and that ERß message is
down-regulated by gonadotropins in granulosa cells, suggesting that the
functional significance of estrogen action in the rat ovary may be
mediated primarily by ERß (18).
The detailed biological significance of the existence of two ERs is presently unclear. Perhaps the existence of two ER subtypes may provide, at least in part, an explanation for the selective actions of estrogens and certain antiestrogens in different target tissues (19, 20).
In this paper we describe the cloning of the mouse ovary ERß-cDNA and
the characterization of the mouse ERß protein with respect to DNA
binding, homo- and heterodimerization, and transactivational
functions. Finally, cotransfection of both ER subtypes with an estrogen
response element (ERE) containing reporter gene construct showed that
the formation of heterodimeric ER/ERß complexes may indeed
constitute a novel estrogenic gene-regulatory pathway.
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RESULTS AND DISCUSSION |
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TOPABSTRACTINTRODUCTIONRESULTS AND DISCUSSIONMATERIALS AND METHODSREFERENCES |
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1.5 kbp) were subcloned and sequenced. The open reading frame
of these clones displayed a high degree of amino acid identity with the
rERß protein and were therefore recognized as the mouse homolog of
rERß and will hereafter be referred to as the mouse ERß (mERß).
As shown in Fig. 1
, the mERß amino acid
sequence also manifests considerable similarities to mouse and rat
ER in the DNA- and ligand-binding domains (Fig. 1). Several amino
acid residues that have been demonstrated to be required for
high-affinity binding of estradiol by ER (21) were found to be
conserved in rat ERß. The rat ERß binds estradiol (E2)
with an affinity very comparable to that of ER (17). Since the same
amino acid residues are also conserved in the ligand-binding domain of
mERß, we concluded that mERß should bind E2 in a
similar manner as the rat ERß.
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(EGCKA) differs from the P-box of
other members of the steroid receptor subgroup, such as glucocorticoid
receptor and progesterone receptor, with the result that ER
recognizes DNA elements contrasting from the sequences recognized by GR
and PR. The consensus ERE consists of a palindromic repeat of the core
sequence AGGTCA spaced by three nucleotides (24). The high degree of
conservation in the DBD of the and ß ER subtypes (96%, Fig. 1
) and the absolute identity of the P-box sequences strongly suggest a
shared DNA recognition specificity between the two ER subtypes. We
consequently performed DNA binding studies with mERß using
radiolabeled consensus ERE oligonucleotides.
Mouse ERß and human ER protein were synthesized in
vitro in a rabbit reticulocyte lysate (RRL) system before
incubation with E2 and a radiolabeled double-stranded ERE
oligonucleotide. The resulting DNA-protein complexes were analyzed by
electrophoretic gel mobility shift assay (Fig. 2). ERß binds to the wild type
consensus ERE both in the absence or presence of E2 (lanes
2 and 3), but not to an ERE mutated in both half-sites (Fig. 2, lanes 7
and 8). This coincides with the binding specificity of ER (Fig. 2, lanes 4 and 5 and lanes 9 and 10, respectively). In contrast to results
obtained in a recent study by Tremblay and co-workers (25), we did not
observe a reduced affinity for the ERE by ERß when equal amounts of
ER and ERß protein were used (as quantitated by
[35S]methionine labeling). The reason for this
discrepancy is unclear.
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). This region of steroid receptors has
not been shown to participate in DNA recognition, and we therefore
anticipated that the truncated mERß would be able to bind to DNA as
efficiently as the wild type receptor. The mERß construct was also
tagged with a nine-amino acid HA1-epitope, which is recognized by the
monoclonal 12CA5 antibody (28). The truncated epitope-tagged mERß
(ERß-TAG) in electrophoretic mobility shift assays gave rise to a
DNA-protein complex clearly distinguishable from the complex formed by
the wild type ERß (ERß-wt), (Fig. 3B). After mixed synthesis of
ERß-TAG and ERß-wt in the RRL system, a third complex of
intermediate mobility was visible (Fig. 3B, lanes 2–4) representing
the dimer between wtERß and ERß-TAG. This experiment clearly shows
that ERß binds to DNA as a homodimer, similar to the binding of ER
(Refs. 1 and 2 and references therein).
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Does Not Inhibit ER-Stimulated
Activity of an ERE-Reporter Gene Construct- or mERß-expressing plasmids, or both. As shown in
Fig. 4, the E2-stimulated
activity of the reporter gene construct by mERß was lower when
compared with the activity obtained with ER when studied under the
same transfection conditions. The 2-fold lower
E2-stimulated activity was not due to squelching of mERß,
since the reporter activity was dose-dependent with regard to the
amount of cotransfected mERß-expression plasmid (data not shown).
Although ERß and ER are highly homologous in the DBD and in parts
of the ligand-binding domain (LBD), there remain substantial
differences, particularly in the N-terminal A/B domain (Fig. 1). This
domain contains the AF-1 transcriptional activity function of ER
(29) and may have a similar function in ERß. The diverging A/B
domains and/or dissimilarities in the LBD of ERß and ER may result
in differences in maximal transactivational activity of both ER
subtypes. The slightly lower maximal transactivational activity of
ERß compared with ER has also been observed by other investigators
(25, 30). When ER and ERß were cotransfected, however, the
reporter activity did not change significantly, when compared with the
activity observed with ER alone (Fig. 4). Based on these
observations we conclude that ERß does not repress the activity of
ER. Furthermore, if the two receptors were competing as homodimers
for the ERE-binding sites, higher concentrations of ERß would be
expected to result in a decrease in reporter activity, toward the
activity pattern of ERß alone. Since no sign of such a competition
was observed, we speculated that an interaction was taking place
between the two ER subtypes (although we cannot rule out that ER
alone is responsible for the transcriptional activity of the reporter
gene). To be able to monitor a possible interaction between ERß and
ER, we used a mammalian cell two-hybrid system.
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in Vivo and in
Vitro, and the VP16
transactivation domain was coupled to full-length mER and mERß,
respectively (Fig. 5A). The chimera
constructs were then cotransfected together with a Gal4-luciferase
reporter construct into COS 7 cells. Comparisons were made between
cells with or without E2 incubation. Reporter activity
remained low when cells were transfected with the vectors expressing
only the Gal4-DBD or the VP16 transactivation domain, or when either of
these constructs was transfected together with any of the hybrid
constructs (Fig. 5B). In contrast, when the Gal4-mER construct was
transfected together with the VP16-mER construct or the VP16-mERß
construct, the activity of the reporter was induced approximately 3- to
4-fold, respectively (Fig. 5B, two most right-hand panels,
white bars), compared with reporter transfected alone,
indicating an interaction between the chimeric proteins. In the
presence of E2 the induction levels rose to about 18-fold
(Fig. 5B, two most right-hand panels, black
bars). These results clearly demonstrate an interaction between
the mERß and mER proteins in vivo, suggesting that the
transcriptional activity observed during coexpression of ER and
ERß (Fig. 4) is, at least in part, due to the formation of
ER/ERß heterodimers. The rise in reporter activity observed,
especially in the presence of E2 (5-fold), with the
Gal4-mER hybrid and the original VP-16 construct (Fig. 5B, second panel) is in all likelihood due to the
ligand-inducible transactivation function of ER itself, when
directed to the promoter of the Gal4 reporter gene construct by binding
via the Gal4 DBD.
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and a
GST-mERß fusion protein, in order to detect a direct interaction
between the two proteins in vitro. As shown in Fig. 5C, ER could be successfully coprecipitated with the GST-mERß fusion
protein but not with the GST alone (compare lane 2 to lane 3, lane
1 = input of ER, 20%), demonstrating a direct interaction
between both ER subtypes.
ERß and ER Form DNA-Binding Heterodimers
The results from the two-hybrid assay and from the pulldown
experiment suggested that ER and ERß are able to form
heterodimers. In combination with the results from the cotransfection
experiments (Fig. 4), it appeared likely that the putative ERß/ER
heterodimer would be able to bind to an ERE.
We performed electrophoretic mobility shift assay with in
vitro synthesized ER and ERß to examine this possibility.
Because the wild type ERß migrates closely with ER on native gels
(see Fig. 2), we decided to use the truncated ERß-TAG (described
above and in Materials and Methods) in order to identify
DNA-protein complexes. ERß-TAG and ER were synthesized in
vitro and then mixed at increasing and decreasing amounts,
respectively, incubated on ice, followed by incubation with the
32P-labeled ERE. An ERE-protein complex of intermediate
mobility was formed in samples in which the two ER subtypes were
coincubated (Fig. 6A, lanes 2–5),
probably representing a heterodimeric complex between ERß-TAG and
ER. This putative heterodimerization was also evident when
full-length wt ERß was used instead of ERß-TAG, but the complexes
were not as easily distinguished (not shown).
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antibody and the 12CA5 antibody. Figure 6B shows the result of
incubation with the monoclonal ER antibody 1D5. The ER homodimer
is efficiently supershifted with this antibody as expected (Fig. 6B, lane 3 compared with lane 2), whereas the ERß homodimer is not
affected (lanes 9 and 10). The intermediate complex formed in the
presence of both ER and ERß-TAG is also supershifted with the
ER antibody (Fig. 6B, lanes 5–8 compared with lane 4), thus
demonstrating the presence of ER within the heterodimeric complex.
In Fig. 6C essentially the same experiment has been repeated using the
12CA5 monoclonal antibody directed against the HA-epitope of
mERß-TAG. The 12CA5 antibody successfully interacts with both the
ERß homodimer and the intermediate complex previously shown to
contain also ER (Fig. 6C, lanes 10 and 4–7). The 12CA5 antibody
does not cross-react with the ER homodimer (lane 2). These results
clearly demonstrate that the intermediate complex that is formed when
ER and ERß are coincubated contains both receptors and is a true
heterodimeric complex.
To determine whether both partners of the heterodimer participate in
DNA binding, experiments were performed with an ERE mutated in one of
the half-sites at a position previously demonstrated to be crucial for
efficient binding by the ER protein (31). In crystallographic
studies of the DBD of ER bound to the ERE (31), it was established
that a mutation in one of the half-sites resulted in reduced
cooperativity in binding to the second half-site, due to lack of proper
interaction between the dimer interfaces present in the DBD. Binding by
ER to such a mutated ERE was therefore less efficient. We found that
ERß did not bind to this mutated ERE in analogy to the ER (Fig. 6D, lanes 7 and 6, respectively). In addition, no protein-DNA complex
was formed with the heterodimer (lane 8), indicating that cooperativity
in DNA binding is also required for efficient DNA binding by
ERß/ER heterodimers.
Furthermore, the ERß as well as ER were unable to bind to
oligonucleotides containing direct repeats of the core sequence AGGTCA
spaced by one or four nucleotides (DR1 or DR4), irrespective of the
presence or absence of retinoid X receptor (data not shown).
Our findings on ER/ERß heterodimerization and the recent
demonstration of GR/MR heterodimerization (32, 33) challenge the
commonly held view that steroid receptors form only homodimers.
Previous biochemical and structural evidence indicated that steroid
receptors form homodimers through a dimerization interface within their
zinc finger DNA binding domain, and a generally much stronger
dimerization interface within the ligand binding domain (Refs. 1 and 22
and references therein). Further studies will be required to localize
the dimerization interfaces involved in the formation of ER/ERß
heterodimers.
The rat tissue distribution and/or relative level of ER and ERß
mRNA seems to be quite different; that is, moderate to high expression
in uterus, testis, pituitary, ovary, kidney, epididymis, and adrenal
for ER and prostate, ovary, uterus, lung, bladder, brain, and testis
for ERß (17). This may imply that in testis and ovary both subtypes
are expressed to some extent. In the mouse, both ER mRNAs can be found
in ovary and uterus (not shown). In the rat, hypothalamus ER and
ERß are coexpressed in certain regions, most likely in the same
neurons (34). The coexpression of ER and ERß in the same tissue
and/or cells suggests the interesting possibility that ER and ERß
proteins may interact with each other. In this study we have indeed
shown that the two ER subtypes have the ability to form heterodimers.
The discovery of an ERß protein and the ability of ER and ERß to
form heterodimers strongly suggest the existence of two previously
unrecognized pathways of estrogen signaling: via ERß homodimers in
cells exclusively expressing this subtype and via ER/ERß
heterodimers in cells expressing both subtypes (Fig. 7). The ERß homodimers and the
ER/ERß heterodimers may possibly interact with novel response
elements, different from the known EREs. By such a mechanism the
physiological regulatory potential of estrogenic hormones may be
greatly expanded. Different target tissues may respond differently to
the same hormonal stimulus due to alternative composition of receptors.
Varying ratios of ER and ERß in different cells, resulting in
different populations of homo- and heterodimers, could constitute a
hitherto unrecognized mechanism involved in the tissue- and cell
type-specific effects of estrogens and certain antiestrogens (19, 20).
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MATERIALS AND METHODS |
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TOPABSTRACTINTRODUCTIONRESULTS AND DISCUSSIONMATERIALS AND METHODSREFERENCES |
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Plasmid Constructs
For in vitro transcription/translation in RRL,
pTKS-mERß was digested with NcoI and EcoRI,
which yielded a fragment encompassing the entire coding sequence (cds),
which was inserted Sp6-sense into
NcoI/EcoRI-digested pSP72 (Promega, Madison, WI),
thus generating pSP72-mERß. pSP72-mERß-TAG was made by replacement
of nucleotides 1–273 of the mERß cds with oligonucleotides
5'-CATGGGCTACCCCTACGACGTGCCCGACTACGCCGTGAACA and
5'-CTAGTGTTCACGGCGTAGTCGGGCACGTCGTAGGGGTAGCC, which encode the HA1
epitope recognized by the 12CA5 monoclonal antibody. The plasmid
pSP72-hER has been described elsewhere (36). For transfections of
mammalian cells the XhoI/BglII-fragment from
pSP72-mERß was inserted into the pSG5 expression vector (Stratagene,
La Jolla, CA) digested with EcoRI/BglII; the
XhoI-site of the mERß-fragment and the
EcoRI-site of pSG5 were filled in with Klenow fragment to
allow blunt-ended ligation. The vector pSG5-hER was made by excising
hER cDNA from pSP72-hER with EcoRI and SacI
and inserting it into pSG5 digested with EcoRI and
BglII. To enable this ligation, the SacI site was
filled in with T4 DNA-polymerase and BglII with Klenow
fragment. The reporter construct 2xERE-TK-Luc was constructed by
subcloning of a tandem ERE (39) with XhoI overhangs into the
SalI site of the p19-TK-Luc reporter plasmid (40). The
Gal4-mER and VP16-mER two-hybrid constructs were made through PCR
amplification of the plasmid MOR101 (containing the mouse ER cDNA)
(41) to introduce a KpnI site upstream of the start codon of
the mER, with the use of oligonucleotides mER-ATG
(5'-GCCAGGTACCATGGCCATGACC) and mER-EagI
(5'-CCCAGGCTGTTGGCACTGAAGGC). The 275-bp long PCR product was cut with
KpnI and EagI, MOR101 was digested with
EagI (nucleotide 460 in the mouse ER cDNA) and
BamHI 3' of the mER cDNA), and both fragments were
subcloned into the KpnI and BamHI sites of
pCMX-Gal4 or pCMX-VP16 (42). VP16-ERß was made by in-frame insertion
of the NcoI/EcoRI fragment of pSP72-mERß into
the EcoRV/EcoRI sites of pCMX-VP16, and the
NcoI site of mERß was filled in with Klenow fragment to
enable the ligation. The Gal4-luciferase reporter construct used in the
two-hybrid assay has been described elsewhere (43). The pGST-mERß was
generated by in-frame ligation of the NcoI/EcoRI
fragment from pSP72-mERß into the corresponding sites of pGST (I.
Pongratz and F. Delauney, unpublished data) creating a GST-mERß
fusion that could be translated in the RRL system.
Cell Culture and Transient Transfections
Cells from the human fetal kidney cell line 293 were routinely
cultured in a 1:1 mixture of Ham’s Nutrient mixture F12 (F12, GIBCO
BRL) and DMEM (GIBCO-BRL) supplemented with 7.5% FBS, 0.5%
nonessential amino acids (NEA, GIBCO BRL) and 1% PEST (100 U
penicillin/ml and 100 µg streptomycin/ml). Cells were seeded in
six-well plates 24 h before transfection. Transfections using the
Lipofectin (GIBCO BRL) reagent were performed as described by the
manufacturer in a serum- and antibiotic-free mixture of 1:1 of F12 and
phenol-red free DMEM with 0.75 µg of the 2xERE-TK-Luc reporter and
0.1–0.4 µg of pSG5-ER or pSG5-ERß as indicated. The pSG5 vector
was used to equalize plasmid concentrations, and 0.1 µg of a
placental alkaline phosphatase (AP) expression plasmid (44) was
included to control for differences in transfection efficiences. Medium
was changed to a phenol red-free mixture of F12 and DMEM containing
7.5% dextran-coated charcoal-treated FBS, 0.5% NEA, and 1% PEST
after 24 h. Hormone or vehicle (0.1% ethanol) was added
simultaneously. Cells were allowed to stand for 48 h with a
renewed change of media and hormone after 24 h. Media were
collected for assaying of AP activity. The cells were harvested in 10
mM Tris-HCl/10 mM EDTA/150 mM NaCl
and centrifuged for 4 min at 4000 rpm, supernatant was removed, and
cell pellets were lysed in Lysis Buffer 2 (Bio-Orbit, Turku, Finland).
Luciferase activity was measured using the GenGlow system (Bio Orbit).
The results are presented as the mean ± SD of fold
induction of three separate transfections performed in duplicate.
COS 7 cells were routinely maintained in DMEM (GIBCO BRL) supplemented
with 5% FBS and 1% PEST. For transient transfections, cells were
seeded in six-well plates 24 h prior to transfection.
Transfections were carried out with Lipofectin reagent in phenol red-
free DMEM without serum and antibiotics, using 0.5 µg of the
GAL4-luciferase reporter construct and 0.1 µg of each of the
two-hybrid expression plasmids as indicated in the legend to Fig. 5B.
Expression vector concentrations were kept constant in all
transfections by addition of the original pCMX-Gal4 or pCMX-VP16
plasmids, and 0.2 µg of the AP expression vector was included in all
transfections as an internal control for transfection efficiency. Cells
were left in the Lipofectin-DNA mixture for 24 h after which the
medium was changed to phenol red-free DMEM supplemented with 5%
dextran-coated charcoal-treated FBS and 1% PEST. Hormone (1
µM E2) or vehicle (0.1% ethanol) was added.
After 24 h cells were harvested as described previously, and
luciferase activity was measured. All samples were normalized against
the activity of the AP internal standard. Transfections were carried
out in duplicate, and the results are presented as fold induction and
represent the mean value ± SD of three separate
experiments.
In Vitro Translation, GST Pulldown, and DNA-Binding
Assays
For the ERE-binding studies, 1 µg pSP72-mERß or 1 µg
pSP72-hER was transcribed/translated in the TNT-coupled RRL system
(Promega) with Sp6 RNA polymerase, according to the manufacturer’s
instructions, in the presence of 100 nM 17ß-estradiol or
vehicle (0.01% ethanol). Five microliters of the lysate were used in
each DNA-binding reaction with a 32P end-labeled wild type
or double-mutated ERE as indicated in the legend to Fig. 2. Protein-DNA
complexes were separated on 5% polyacrylamide/0.25x Tris-borate-EDTA
gels at 10 V/cm, followed by drying and autoradiography at -70
C.
In the homodimerization experiments, increasing amounts of pSP72-mERß-TAG (0, 0.1, 0.2, 0.3, and 0.4 µg) were translated in the RRL together with decreasing amounts of pSP72-mERß (0.4, 0.3, 0.2, 0.1, and 0 µg). Five microliters of programmed lysate were used in each DNA-binding reaction with radiolabeled ERE.
For the heterodimerization studies, 1 µg pSP72-hER or pSP72-mERß
was translated in RRL. In the experiment of Fig. 6A, 5, 4, 3, 2, 1 or 0
µl of ER-containing lysate were mixed with 0, 1, 2, 3, 4, or 5
µl of ERß lysate and incubated 15 min on ice before the DNA-binding
reaction with radiolabeled ERE.
Translations were carried out in the same manner for the
antibody-upshift experiments, and 4 µl ER or ERß lysate,
respectively, or a mixture of 2 µl of each was incubated for 15 min
on ice. Thereafter, 1.5 µl monoclonal ER antibody 1D5 (Dako,
Carpinteria, CA) or 12CA5 TAG antibody (BAbCOf) were added to the
respective homodimers and 0.5, 1, 2, or 3 µl of each antibody were
added to the heterodimer reactions. The DNA-binding reaction was
started immediately. The single-mutated ERE used in the band shift
assay in Fig. 6D has been described previously (36). Four microliters
of ER or ERß protein containing RRL or a mix of 2 µl of each
were used in the DNA-binding assay.
For the GST pulldown experiments, pSP72-hER was translated in the
presence of [35S]methionine in RRL and pGST-mERß or the
original pGST was translated in RRL in the absence of radiolabeled
amino acids. Five microliters of ER-containing lysate were mixed
with 5 µl lysate containing GST-mERß or GST-protein. Samples were
incubated for 15 min on ice before 50 µl GST-Sepharose diluted in PBS
were added to each sample followed by 30 min of incubation on ice. The
Sepharose beads were washed four times in PBS/0.1% Triton X-100, and
bound proteins were eluted by incubation in 2x SDS-buffer for 5 min at
100 C. ER lysate (=20%) was loaded as input together with the
eluted samples on a 10% SDS-PAGE and run at 150 V. The gel was
immersed in 1 M salicylic acid for 20 min, dried, and
autoradiographed at -70 C.
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ACKNOWLEDGMENTS |
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FOOTNOTES |
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This work was supported by a grant from the Swedish Cancer Society to J-ÅG and GGJMK was supported by a visiting scientist fellowship from the Karolinska Institute.
The sequence reported in this paper has been deposited in the GenBank database (AJ000220).
Received for publication December 18, 1996. Revision received May 7, 1997. Accepted for publication June 2, 1997.
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REFERENCES |
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TOPABSTRACTINTRODUCTIONRESULTS AND DISCUSSIONMATERIALS AND METHODSREFERENCES |
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 S.-H. Yang, S. N. Sarkar, R. Liu, E. J. Perez, X. Wang, Y. Wen, L.-J. Yan, and J. W. Simpkins Estrogen Receptor {beta} as a Mitochondrial Vulnerability Factor J. Biol. Chem., April 3, 2009; 284(14): 9540 - 9548. [Abstract] [Full Text] [PDF]
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 M. Brown, J. Ning, J. A. Ferreira, J. L. Bogener, and D. B. Lubahn Estrogen receptor-{alpha} and -{beta} and aromatase knockout effects on lower limb muscle mass and contractile function in female mice Am J Physiol Endocrinol Metab, April 1, 2009; 296(4): E854 - E861. [Abstract] [Full Text] [PDF]
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 A. Cignarella, C. Bolego, V. Pelosi, C. Meda, A. Krust, C. Pinna, R. M. Gaion, E. Vegeto, and A. Maggi Distinct Roles of Estrogen Receptor-{alpha} and {beta} in the Modulation of Vascular Inducible Nitric-Oxide Synthase in Diabetes J. Pharmacol. Exp. Ther., January 1, 2009; 328(1): 174 - 182. [Abstract] [Full Text] [PDF]
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A. M Davis, J. Mao, B. Naz, J. A Kohl, and C. S Rosenfeld Comparative effects of estradiol, methyl-piperidino-pyrazole, raloxifene, and ICI 182 780 on gene expression in the murine uterus J. Mol. Endocrinol., October 1, 2008; 41(4): 205 - 217. [Abstract] [Full Text] [PDF]
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K. L. Gonzales, M. J. Tetel, and C. K. Wagner Estrogen Receptor (ER) {beta} Modulates ER{alpha} Responses to Estrogens in the Developing Rat Ventromedial Nucleus of the Hypothalamus Endocrinology, September 1, 2008; 149(9): 4615 - 4621. [Abstract] [Full Text] [PDF]
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 A. Orgaard and L. Jensen The Effects of Soy Isoflavones on Obesity Experimental Biology and Medicine, September 1, 2008; 233(9): 1066 - 1080. [Abstract] [Full Text] [PDF]
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 J. Ruegg, E. Swedenborg, D. Wahlstrom, A. Escande, P. Balaguer, K. Pettersson, and I. Pongratz The Transcription Factor Aryl Hydrocarbon Receptor Nuclear Translocator Functions as an Estrogen Receptor {beta}-Selective Coactivator, and Its Recruitment to Alternative Pathways Mediates Antiestrogenic Effects of Dioxin Mol. Endocrinol., February 1, 2008; 22(2): 304 - 316. [Abstract] [Full Text] [PDF]
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 X. Fan, H.-J. Kim, M. Warner, and J.-A. Gustafsson Estrogen receptor beta is essential for sprouting of nociceptive primary afferents and for morphogenesis and maintenance of the dorsal horn interneurons PNAS, August 21, 2007; 104(34): 13696 - 13701. [Abstract] [Full Text] [PDF]
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 S. K. Gruvberger-Saal, P.-O. Bendahl, L. H. Saal, M. Laakso, C. Hegardt, P. Eden, C. Peterson, P. Malmstrom, J. Isola, A. Borg, et al. Estrogen Receptor {beta} Expression Is Associated with Tamoxifen Response in ER{alpha}-Negative Breast Carcinoma Clin. Cancer Res., April 1, 2007; 13(7): 1987 - 1994. [Abstract] [Full Text] [PDF]
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 P. Galluzzo, F. Caiazza, S. Moreno, and M. Marino Role of ER{beta} palmitoylation in the inhibition of human colon cancer cell proliferation Endocr. Relat. Cancer, March 1, 2007; 14(1): 153 - 167. [Abstract] [Full Text] [PDF]
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D. Titolo, F. Cai, and D. D. Belsham Coordinate Regulation of Neuropeptide Y and Agouti-Related Peptide Gene Expression by Estrogen Depends on the Ratio of Estrogen Receptor (ER) {alpha} to ER{beta} in Clonal Hypothalamic Neurons Mol. Endocrinol., September 1, 2006; 20(9): 2080 - 2092. [Abstract] [Full Text] [PDF]
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 M. F. Kritzer Regional, Laminar and Cellular Distribution of Immunoreactivity for ER{beta} in the Cerebral Cortex of Hormonally Intact, Postnatally Developing Male and Female Rats Cereb Cortex, August 1, 2006; 16(8): 1181 - 1192. [Abstract] [Full Text] [PDF]
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 G. Hasenbrink, A. Sievernich, L. Wildt, J. Ludwig, and H. Lichtenberg-Frate Estrogenic effects of natural and synthetic compounds including tibolone assessed in Saccharomyces cerevisiae expressing the human estrogen {alpha} and {beta} receptors FASEB J, July 1, 2006; 20(9): 1552 - 1554. [Abstract] [Full Text] [PDF]
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Y. Yamamoto, R. Moore, H. A. Hess, G. L. Guo, F. J. Gonzalez, K. S. Korach, R. R. Maronpot, and M. Negishi Estrogen Receptor {alpha} Mediates 17{alpha}-Ethynylestradiol Causing Hepatotoxicity J. Biol. Chem., June 16, 2006; 281(24): 16625 - 16631. [Abstract] [Full Text] [PDF]
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 M. R. Meyer, E. Haas, and M. Barton Gender Differences of Cardiovascular Disease: New Perspectives for Estrogen Receptor Signaling Hypertension, June 1, 2006; 47(6): 1019 - 1026. [Full Text] [PDF]
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M. Singh, J. A. Dykens, and J. W. Simpkins Novel Mechanisms for Estrogen-Induced Neuroprotection. Experimental Biology and Medicine, May 1, 2006; 231(5): 514 - 521. [Abstract] [Full Text] [PDF]
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 H. N. Jabbour, R. W. Kelly, H. M. Fraser, and H. O. D. Critchley Endocrine Regulation of Menstruation Endocr. Rev., February 1, 2006; 27(1): 17 - 46. [Abstract] [Full Text] [PDF]
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C. Bodo, A. E. Kudwa, and E. F. Rissman Both Estrogen Receptor-{alpha} and -{beta} Are Required for Sexual Differentiation of the Anteroventral Periventricular Area in Mice Endocrinology, January 1, 2006; 147(1): 415 - 420. [Abstract] [Full Text] [PDF]
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P. Ciana, A. Brena, P. Sparaciari, E. Bonetti, D. Di Lorenzo, and A. Maggi Estrogenic Activities in Rodent Estrogen-Free Diets Endocrinology, December 1, 2005; 146(12): 5144 - 5150. [Abstract] [Full Text] [PDF]
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L. A. Gilad, T. Bresler, J. Gnainsky, P. Smirnoff, and B. Schwartz Regulation of vitamin D receptor expression via estrogen-induced activation of the ERK 1/2 signaling pathway in colon and breast cancer cells J. Endocrinol., June 1, 2005; 185(3): 577 - 592. [Abstract] [Full Text] [PDF]
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 M.B Hawkins, J Godwin, D Crews, and P Thomas The distributions of the duplicate oestrogen receptors ER-{beta}a and ER-{beta}b in the forebrain of the Atlantic croaker (Micropogonias undulatus): evidence for subfunctionalization after gene duplication Proc R Soc B, March 22, 2005; 272(1563): 633 - 641. [Abstract] [Full Text] [PDF]
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 S. H. Kim, A. Tamrazi, K. E. Carlson, and J. A. Katzenellenbogen A Proteomic Microarray Approach for Exploring Ligand-initiated Nuclear Hormone Receptor Pharmacology, Receptor Selectivity, and Heterodimer Functionality Mol. Cell. Proteomics, March 1, 2005; 4(3): 267 - 277. [Abstract] [Full Text] [PDF]
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T. A. Hopp, H. L. Weiss, I. S. Parra, Y. Cui, C. K. Osborne, and S. A. W. Fuqua Low Levels of Estrogen Receptor {beta} Protein Predict Resistance to Tamoxifen Therapy in Breast Cancer Clin. Cancer Res., November 15, 2004; 10(22): 7490 - 7499. [Abstract] [Full Text] [PDF]
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 C. Zhao, L. Xu, M. Otsuki, G. Toresson, K. Koehler, Q. Pan-Hammarstrom, L. Hammarstrom, S. Nilsson, J.-A. Gustafsson, and K. Dahlman-Wright Identification of a functional variant of estrogen receptor beta in an African population Carcinogenesis, November 1, 2004; 25(11): 2067 - 2073. [Abstract] [Full Text] [PDF]
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B Horard, A Castet, P-L Bardet, V Laudet, V Cavailles, and J-M Vanacker Dimerization is required for transactivation by estrogen-receptor-related (ERR) orphan receptors: evidence from amphioxus ERR J. Mol. Endocrinol., October 1, 2004; 33(2): 493 - 509. [Abstract] [Full Text] [PDF]
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A. E. Kudwa, J.-A. Gustafsson, and E. F. Rissman Estrogen Receptor {beta} Modulates Estradiol Induction of Progestin Receptor Immunoreactivity in Male, But Not in Female, Mouse Medial Preoptic Area Endocrinology, October 1, 2004; 145(10): 4500 - 4506. [Abstract] [Full Text] [PDF]
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A.G.B. Hurst, D.W. Goad, M. Mohan, and J.R. Malayer Independent Downstream Gene Expression Profiles in the Presence of Estrogen Receptor {alpha} or {beta} Biol Reprod, October 1, 2004; 71(4): 1252 - 1261. [Abstract] [Full Text] [PDF]
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A Bardin, N Boulle, G Lazennec, F Vignon, and P Pujol Loss of ER{beta} expression as a common step in estrogen-dependent tumor progression Endocr. Relat. Cancer, September 1, 2004; 11(3): 537 - 551. [Abstract] [Full Text] [PDF]
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R. O'Lone, M. C. Frith, E. K. Karlsson, and U. Hansen Genomic Targets of Nuclear Estrogen Receptors Mol. Endocrinol., August 1, 2004; 18(8): 1859 - 1875. [Abstract] [Full Text] [PDF]
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 J.-x. Zhang, D. C. Labaree, G. Mor, and R. B. Hochberg Estrogen to Antiestrogen with a Single Methylene Group Resulting in an Unusual Steroidal Selective Estrogen Receptor Modulator J. Clin. Endocrinol. Metab., July 1, 2004; 89(7): 3527 - 3535. [Abstract] [Full Text] [PDF]
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S.-H. Yang, R. Liu, E. J. Perez, Y. Wen, S. M. Stevens Jr., T. Valencia, A.-M. Brun-Zinkernagel, L. Prokai, Y. Will, J. Dykens, et al. Mitochondrial localization of estrogen receptor {beta} PNAS, March 23, 2004; 101(12): 4130 - 4135. [Abstract] [Full Text] [PDF]
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R. L. Arnett-Mansfield, A. deFazio, P. A. Mote, and C. L. Clarke Subnuclear Distribution of Progesterone Receptors A and B in Normal and Malignant Endometrium J. Clin. Endocrinol. Metab., March 1, 2004; 89(3): 1429 - 1442. [Abstract] [Full Text] [PDF]
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U. I. L. Modder, A. Sanyal, A. E. Kearns, J. D. Sibonga, E. Nishihara, J. Xu, B. W. O'Malley, E. L. Ritman, B. L. Riggs, T. C. Spelsberg, et al. Effects of Loss of Steroid Receptor Coactivator-1 on the Skeletal Response to Estrogen in Mice Endocrinology, February 1, 2004; 145(2): 913 - 921. [Abstract] [Full Text] [PDF]
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 S. Paruthiyil, H. Parmar, V. Kerekatte, G. R. Cunha, G. L. Firestone, and D. C. Leitman Estrogen Receptor {beta} Inhibits Human Breast Cancer Cell Proliferation and Tumor Formation by Causing a G2 Cell Cycle Arrest Cancer Res., January 1, 2004; 64(1): 423 - 428. [Abstract] [Full Text] [PDF]
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 C. Patrone, T. N. Cassel, K. Pettersson, Y.-S. Piao, G. Cheng, P. Ciana, A. Maggi, M. Warner, J.-A. Gustafsson, and M. Nord Regulation of Postnatal Lung Development and Homeostasis by Estrogen Receptor {beta} Mol. Cell. Biol., December 1, 2003; 23(23): 8542 - 8552. [Abstract] [Full Text] [PDF]
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C. K. Cheng, B. K. C. Chow, and P. C. K. Leung An Activator Protein 1-Like Motif Mediates 17{beta}-Estradiol Repression of Gonadotropin-Releasing Hormone Receptor Promoter via an Estrogen Receptor {alpha}-Dependent Mechanism in Ovarian and Breast Cancer Cells Mol. Endocrinol., December 1, 2003; 17(12): 2613 - 2629. [Abstract] [Full Text] [PDF]
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Y. Ikeda, A. Nagai, M.-A. Ikeda, and S. Hayashi Sexually Dimorphic and Estrogen-Dependent Expression of Estrogen Receptor {beta} in the Ventromedial Hypothalamus during Rat Postnatal Development Endocrinology, November 1, 2003; 144(11): 5098 - 5104. [Abstract] [Full Text] [PDF]
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M. Otsuki, H. Gao, K. Dahlman-Wright, C. Ohlsson, N. Eguchi, Y. Urade, and J.-A. Gustafsson Specific Regulation of Lipocalin-Type Prostaglandin D Synthase in Mouse Heart by Estrogen Receptor {beta} Mol. Endocrinol., September 1, 2003; 17(9): 1844 - 1855. [Abstract] [Full Text] [PDF]
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 J. Matthews and J.-A. Gustafsson Estrogen Signaling: A Subtle Balance Between ER{alpha} and ER{beta} Mol. Interv., August 1, 2003; 3(5): 281 - 292. [Abstract] [Full Text] [PDF]
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J. Frasor, D. H. Barnett, J. M. Danes, R. Hess, A. F. Parlow, and B. S. Katzenellenbogen Response-Specific and Ligand Dose-Dependent Modulation of Estrogen Receptor (ER) {alpha} Activity by ER{beta} in the Uterus Endocrinology, July 1, 2003; 144(7): 3159 - 3166. [Abstract] [Full Text] [PDF]
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J. F. Couse, M. M. Yates, V. R. Walker, and K. S. Korach Characterization of the Hypothalamic-Pituitary-Gonadal Axis in Estrogen Receptor (ER) Null Mice Reveals Hypergonadism and Endocrine Sex Reversal in Females Lacking ER{alpha} But Not ER{beta} Mol. Endocrinol., June 1, 2003; 17(6): 1039 - 1053. [Abstract] [Full Text] [PDF]
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S. A. W. Fuqua, R. Schiff, I. Parra, J. T. Moore, S. K. Mohsin, C. K. Osborne, G. M. Clark, and D. C. Allred Estrogen Receptor {beta} Protein in Human Breast Cancer: Correlation with Clinical Tumor Parameters Cancer Res., May 15, 2003; 63(10): 2434 - 2439. [Abstract] [Full Text] [PDF]
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S. W. Mitra, E. Hoskin, J. Yudkovitz, L. Pear, H. A. Wilkinson, S. Hayashi, D. W. Pfaff, S. Ogawa, S. P. Rohrer, J. M. Schaeffer, et al. Immunolocalization of Estrogen Receptor {beta} in the Mouse Brain: Comparison with Estrogen Receptor {alpha} Endocrinology, May 1, 2003; 144(5): 2055 - 2067. [Abstract] [Full Text] [PDF]
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Y. Bai and V. Giguere Isoform-Selective Interactions between Estrogen Receptors and Steroid Receptor Coactivators Promoted by Estradiol and ErbB-2 Signaling in Living Cells Mol. Endocrinol., April 1, 2003; 17(4): 589 - 599. [Abstract] [Full Text] [PDF]
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M. K. Lindberg, S. Moverare, S. Skrtic, H. Gao, K. Dahlman-Wright, J.-A. Gustafsson, and C. Ohlsson Estrogen Receptor (ER)-{beta} Reduces ER{alpha}-Regulated Gene Transcription, Supporting a "Ying Yang" Relationship between ER{alpha} and ER{beta} in Mice Mol. Endocrinol., February 1, 2003; 17(2): 203 - 208. [Abstract] [Full Text] [PDF]
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T. A. Henderson, P. T. K. Saunders, A. Moffett-King, N. P. Groome, and H. O. D. Critchley Steroid Receptor Expression in Uterine Natural Killer Cells J. Clin. Endocrinol. Metab., January 1, 2003; 88(1): 440 - 449. [Abstract] [Full Text] [PDF]
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A. J. Jakimiuk, S. R. Weitsman, H.-W. Yen, M. Bogusiewicz, and D. A. Magoffin Estrogen Receptor {alpha} and {beta} Expression in Theca and Granulosa Cells from Women with Polycystic Ovary Syndrome J. Clin. Endocrinol. Metab., December 1, 2002; 87(12): 5532 - 5538. [Abstract] [Full Text] [PDF]
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J. B. Hodgin and N. Maeda Minireview: Estrogen and Mouse Models of Atherosclerosis Endocrinology, December 1, 2002; 143(12): 4495 - 4501. [Abstract] [Full Text] [PDF]
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S. B. Pillai, J. M. Jones, and R. D. Koos Treatment of Rats with 17{beta}-Estradiol or Relaxin Rapidly Inhibits Uterine Estrogen Receptor {beta}1 and {beta}2 Messenger Ribonucleic Acid Levels Biol Reprod, December 1, 2002; 67(6): 1919 - 1926. [Abstract] [Full Text] [PDF]
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H. O. D. Critchley, T. A. Henderson, R. W. Kelly, G. S. Scobie, L. R. Evans, N. P. Groome, and P. T. K. Saunders Wild-Type Estrogen Receptor (ER{beta}1) and the Splice Variant (ER{beta}cx/{beta}2) Are Both Expressed within the Human Endometrium throughout the Normal Menstrual Cycle J. Clin. Endocrinol. Metab., November 1, 2002; 87(11): 5265 - 5273. [Abstract] [Full Text] [PDF]
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D. A. Schreihofer, D. F. Rowe, E. F. Rissman, E. M. Scordalakes, J.-a. Gustafsson, and M. A. Shupnik Estrogen Receptor-{alpha} (ER{alpha}), But Not ER{beta}, Modulates Estrogen Stimulation of the ER{alpha}-Truncated Variant, TERP-1 Endocrinology, November 1, 2002; 143(11): 4196 - 4202. [Abstract] [Full Text] [PDF]
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 N. Vasudevan, S. Ogawa, and D. Pfaff Estrogen and Thyroid Hormone Receptor Interactions: Physiological Flexibility by Molecular Specificity Physiol Rev, October 1, 2002; 82(4): 923 - 944. [Abstract] [Full Text] [PDF]
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K.-i. Matsuda, I. Ochiai, M. Nishi, and M. Kawata Colocalization and Ligand-Dependent Discrete Distribution of the Estrogen Receptor (ER){alpha} and ER{beta} Mol. Endocrinol., October 1, 2002; 16(10): 2215 - 2230. [Abstract] [Full Text] [PDF]
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W. N. Jefferson, J. F. Couse, E. Padilla-Banks, K. S. Korach, and R. R. Newbold Neonatal Exposure to Genistein Induces Estrogen Receptor (ER){alpha} Expression and Multioocyte Follicles in the Maturing Mouse Ovary: Evidence for ER{beta}-Mediated and Nonestrogenic Actions Biol Reprod, October 1, 2002; 67(4): 1285 - 1296. [Abstract] [Full Text] [PDF]
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H. Buteau-Lozano, M. Ancelin, B. Lardeux, J. Milanini, and M. Perrot-Applanat Transcriptional Regulation of Vascular Endothelial Growth Factor by Estradiol and Tamoxifen in Breast Cancer Cells: A Complex Interplay between Estrogen Receptors {alpha} and {beta} Cancer Res., September 1, 2002; 62(17): 4977 - 4984. [Abstract] [Full Text] [PDF]
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X. Ni, R. C. Nicholson, B. R. King, E.-C. Chan, M. A. Read, and R. Smith Estrogen Represses whereas the Estrogen-Antagonist ICI 182780 Stimulates Placental CRH Gene Expression J. Clin. Endocrinol. Metab., August 1, 2002; 87(8): 3774 - 3778. [Abstract] [Full Text] [PDF]
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H.-J. Huang, J. D. Norris, and D. P. McDonnell Identification of a Negative Regulatory Surface within Estrogen Receptor {alpha} Provides Evidence in Support of a Role for Corepressors in Regulating Cellular Responses to Agonists and Antagonists Mol. Endocrinol., August 1, 2002; 16(8): 1778 - 1792. [Abstract] [Full Text] [PDF]
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M.-M. Liu, C. Albanese, C. M. Anderson, K. Hilty, P. Webb, R. M. Uht, R. H. Price Jr., R. G. Pestell, and P. J. Kushner Opposing Action of Estrogen Receptors alpha and beta on Cyclin D1 Gene Expression J. Biol. Chem., June 28, 2002; 277(27): 24353 - 24360. [Abstract] [Full Text] [PDF]
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B. L. Riggs, S. Khosla, and L. J. Melton III Sex Steroids and the Construction and Conservation of the Adult Skeleton Endocr. Rev., June 1, 2002; 23(3): 279 - 302. [Abstract] [Full Text] [PDF]
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P. T. K. Saunders, M. R. Millar, S. Macpherson, D. S. Irvine, N. P. Groome, L. R. Evans, R. M. Sharpe, and G. A. Scobie ER{beta}1 and the ER{beta}2 Splice Variant (ER{beta}cx/{beta}2) Are Expressed in Distinct Cell Populations in the Adult Human Testis J. Clin. Endocrinol. Metab., June 1, 2002; 87(6): 2706 - 2715. [Abstract] [Full Text] [PDF]
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D. Lopez, M. D. Sanchez, W. Shea-Eaton, and M. P. McLean Estrogen Activates the High-Density Lipoprotein Receptor Gene via Binding to Estrogen Response Elements and Interaction with Sterol Regulatory Element Binding Protein-1A Endocrinology, June 1, 2002; 143(6): 2155 - 2168. [Abstract] [Full Text] [PDF]
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A. Menuet, E. Pellegrini, I. Anglade, O. Blaise, V. Laudet, O. Kah, and F. Pakdel Molecular Characterization of Three Estrogen Receptor Forms in Zebrafish: Binding Characteristics, Transactivation Properties, and Tissue Distributions Biol Reprod, June 1, 2002; 66(6): 1881 - 1892. [Abstract] [Full Text] [PDF]
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G. J. Pepe, R. B. Billiar, M. G. Leavitt, N. C. Zachos, J. A. Gustafsson, and E. D. Albrecht Expression of Estrogen Receptors {alpha} and {beta} in the Baboon Fetal Ovary Biol Reprod, April 1, 2002; 66(4): 1054 - 1060. [Abstract] [Full Text] [PDF]
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R. Nie, Q. Zhou, E. Jassim, P. T.K. Saunders, and R. A. Hess Differential Expression of Estrogen Receptors {alpha} and {beta} in the Reproductive Tractsof Adult Male Dogs and Cats Biol Reprod, April 1, 2002; 66(4): 1161 - 1168. [Abstract] [Full Text] [PDF]
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E. F. Rissman, A. L. Heck, J. E. Leonard, M. A. Shupnik, and J.-A. Gustafsson Disruption of estrogen receptor beta gene impairs spatial learning in female mice PNAS, March 19, 2002; 99(6): 3996 - 4001. [Abstract] [Full Text] [PDF]
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 T. V. Pham and M. R. Rosen Sex, hormones, and repolarization Cardiovasc Res, February 15, 2002; 53(3): 740 - 751. [Abstract] [Full Text] [PDF]
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C. Orikasa, Y. Kondo, S. Hayashi, B. S. McEwen, and Y. Sakuma Sexually dimorphic expression of estrogen receptor beta in the anteroventral periventricular nucleus of the rat preoptic area: Implication in luteinizing hormone surge PNAS, February 14, 2002; (2002) 52707299. [Abstract] [Full Text] [PDF]
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M.F. Kritzer Regional, Laminar, and Cellular Distribution of Immunoreactivity for ER{alpha} and ER{beta} in the Cerebral Cortex of Hormonally Intact, Adult Male and Female Rats Cereb Cortex, February 1, 2002; 12(2): 116 - 128. [Abstract] [Full Text] [PDF]
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 C. J. Gruber, W. Tschugguel, C. Schneeberger, and J. C. Huber Production and Actions of Estrogens N. Engl. J. Med., January 31, 2002; 346(5): 340 - 352. [Full Text] [PDF]
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 B. McEwen Estrogen Actions Throughout the Brain Recent Prog. Horm. Res., January 1, 2002; 57(1): 357 - 384. [Abstract] [Full Text] [PDF]
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 B. S. McEwen Genome and Hormones: Gender Differences in Physiology: Invited Review: Estrogens effects on the brain: multiple sites and molecular mechanisms J Appl Physiol, December 1, 2001; 91(6): 2785 - 2801. [Abstract] [Full Text] [PDF]
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B. Greco, E. A. Allegretto, M. J. Tetel, and J. D. Blaustein Coexpression of ER{beta} with ER{alpha} and Progestin Receptor Proteins in the Female Rat Forebrain: Effects of Estradiol Treatment Endocrinology, December 1, 2001; 142(12): 5172 - 5181. [Abstract] [Full Text] [PDF]
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A. Warnmark, A. Wikstrom, A. P. H. Wright, J.-A. Gustafsson, and T. Hard The N-terminal Regions of Estrogen Receptor alpha and beta Are Unstructured in Vitro and Show Different TBP Binding Properties J. Biol. Chem., November 30, 2001; 276(49): 45939 - 45944. [Abstract] [Full Text] [PDF]
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M. H. Faulds, K. Pettersson, J.-A. Gustafsson, and L.-A. Haldosen Cross-Talk Between ERs and Signal Transducer and Activator of Transcription 5 Is E2 Dependent and Involves Two Functionally Separate Mechanisms Mol. Endocrinol., November 1, 2001; 15(11): 1929 - 1940. [Abstract] [Full Text] [PDF]
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E. R. Levin Genome and Hormones: Gender Differences in Physiology: Invited Review: Cell localization, physiology, and nongenomic actions of estrogen receptors J Appl Physiol, October 1, 2001; 91(4): 1860 - 1867. [Abstract] [Full Text] [PDF]
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D. Lu, Y. Kiriyama, K. Y. Lee, and V. Giguere Transcriptional Regulation of the Estrogen-inducible pS2 Breast Cancer Marker Gene by the ERR Family of Orphan Nuclear Receptors Cancer Res., September 1, 2001; 61(18): 6755 - 6761. [Abstract] [Full Text] [PDF]
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 C. M. Klinge Estrogen receptor interaction with estrogen response elements Nucleic Acids Res., July 15, 2001; 29(14): 2905 - 2919. [Abstract] [Full Text] [PDF]
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J. R. Wood, V. S. Likhite, M. A. Loven, and A. M. Nardulli Allosteric Modulation of Estrogen Receptor Conformation by Different Estrogen Response Elements Mol. Endocrinol., July 1, 2001; 15(7): 1114 - 1126. [Abstract] [Full Text] [PDF]
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H. Cardenas, K.A. Burke, R.M. Bigsby, W.F. Pope, and K.P. Nephew Estrogen Receptor {beta} in the Sheep Ovary During the Estrous Cycle and Early Pregnancy Biol Reprod, July 1, 2001; 65(1): 128 - 134. [Abstract] [Full Text] [PDF]
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S. F. Palter, A. B. Tavares, A. Hourvitz, J. D. Veldhuis, and E. Y. Adashi Are Estrogens of Import to Primate/Human Ovarian Folliculogenesis? Endocr. Rev., June 1, 2001; 22(3): 389 - 424. [Abstract] [Full Text] [PDF]
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C. Pasqualini, D. Guivarc'h, J.-V. Barnier, B. Guibert, J.-D. Vincent, and P. Vernier Differential Subcellular Distribution and Transcriptional Activity of {{Sigma}}E3, {{Sigma}}E4, and {{Sigma}}E3-4 Isoforms of the Rat Estrogen Receptor-{{alpha}} Mol. Endocrinol., June 1, 2001; 15(6): 894 - 908. [Abstract] [Full Text] [PDF]
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 L. J. Havrilesky, C. P. McMahon, E. K. Lobenhofer, R. Whitaker, J. R. Marks, and A. Berchuck Relationship Between Expression of Coactivators and Corepressors of Hormone Receptors and Resistance of Ovarian Cancers to Growth Regulation by Steroid Hormones Reproductive Sciences, April 1, 2001; 8(2): 104 - 113. [Abstract] [PDF]
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M. E. Quaedackers, C. E. Van Den Brink, S. Wissink, R. H. M. M. Schreurs, J.-A. Gustafsson, P. T. Van Der Saag, and B. Van Der Burg 4-Hydroxytamoxifen Trans-Represses Nuclear Factor-{{kappa}}B Activity in Human Osteoblastic U2-OS Cells through Estrogen Receptor (ER){{alpha}}, and Not through ER{{beta}} Endocrinology, March 1, 2001; 142(3): 1156 - 1166. [Abstract] [Full Text] [PDF]
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K. Yokogawa, K. Miya, T. Sekido, Y. Higashi, M. Nomura, R. Fujisawa, K. Morito, Y. Masamune, Y. Waki, S. Kasugai, et al. Selective Delivery of Estradiol to Bone by Aspartic Acid Oligopeptide and Its Effects on Ovariectomized Mice Endocrinology, March 1, 2001; 142(3): 1228 - 1233. [Abstract] [Full Text] [PDF]
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N. Kirma, K. Gill, U. Mandava, and R. R. Tekmal Overexpression of Aromatase Leads to Hyperplasia and Changes in the Expression of Genes Involved in Apoptosis, Cell Cycle, Growth, and Tumor Suppressor Functions in the Mammary Glands of Transgenic Mice Cancer Res., March 1, 2001; 61(5): 1910 - 1918. [Abstract] [Full Text]
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 R. Clarke, F. Leonessa, J. N. Welch, and T. C. Skaar Cellular and Molecular Pharmacology of Antiestrogen Action and Resistance Pharmacol. Rev., March 1, 2001; 53(1): 25 - 72. [Abstract] [Full Text] [PDF]
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J. Kamegai, H. Tamura, T. Shimizu, S. Ishii, H. Sugihara, and I. Wakabayashi Estrogen Receptor (ER){{alpha}}, But Not ER{beta}, Gene Is Expressed in Growth Hormone-Releasing Hormone Neurons of the Male Rat Hypothalamus Endocrinology, February 1, 2001; 142(2): 538 - 543. [Abstract] [Full Text] [PDF]
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S. K. Kang, K.-C. Choi, C.-J. Tai, N. Auersperg, and P. C. K. Leung Estradiol Regulates Gonadotropin-Releasing Hormone (GnRH) and its Receptor Gene Expression and Antagonizes the Growth Inhibitory Effects of GnRH in Human Ovarian Surface Epithelial and Ovarian Cancer Cells Endocrinology, February 1, 2001; 142(2): 580 - 588. [Abstract] [Full Text] [PDF]
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H. A. Harris, R. A. Henderson, R. A. Bhat, and B. S. Komm Regulation of Metallothionein II Messenger Ribonucleic Acid Measures Exogenous Estrogen Receptor-{beta} Activity in SAOS-2 and LNCaPLN3 Cells Endocrinology, February 1, 2001; 142(2): 645 - 652. [Abstract] [Full Text] [PDF]
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 M. POTIER, S. J. ELLIOT, I. TACK, O. LENZ, G. E. STRIKER, L. J. STRIKER, and M. KARL Expression and Regulation of Estrogen Receptors in Mesangial Cells: Influence on Matrix Metalloproteinase-9 J. Am. Soc. Nephrol., February 1, 2001; 12(2): 241 - 251. [Abstract] [Full Text]
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E. D. Lephart, S. B. Call, R. W. Rhees, N. A. Jacobson, K. Scott Weber, J. Bledsoe, and C. Teuscher Neuroendocrine Regulation of Sexually Dimorphic Brain Structure and Associated Sexual Behavior in Male Rats Is Genetically Controlled Biol Reprod, February 1, 2001; 64(2): 571 - 578. [Abstract] [Full Text]
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J. E. Compston Sex Steroids and Bone Physiol Rev, January 1, 2001; 81(1): 419 - 447. [Abstract] [Full Text] [PDF]
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T. Kurita, K.-j. Lee, P. T.K. Saunders, P. S. Cooke, J. A. Taylor, D. B. Lubahn, C. Zhao, S. Mäkelä, J.-A. Gustafsson, R. Dahiya, et al. Regulation of Progesterone Receptors and Decidualization in Uterine Stroma of the Estrogen Receptor-{{alpha}} Knockout Mouse Biol Reprod, January 1, 2001; 64(1): 272 - 283. [Abstract] [Full Text]
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